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. 2011 Oct 27;118(17):4630-4.
doi: 10.1182/blood-2011-01-332049. Epub 2011 Sep 8.

A novel pro-lymphangiogenic function for Th17/IL-17

Sunil K Chauhan  1 , Yiping Jin, Sunali Goyal, Hyun Soo Lee, Thomas A Fuchsluger, Hyung Keun Lee, Reza Dana
Affiliations

A novel pro-lymphangiogenic function for Th17/IL-17

Sunil K Chauhan et al. Blood. .

Abstract

Th17 cells, in addition to their proinflammatory functions, have been recognized as potent inducers of angiogenesis in autoimmune diseases and malignancies. In the present study, we demonstrate distinct mechanisms by which IL-17 induces lymphangiogenesis. Using the mouse cornea micropocket and cell culture assays, our data demonstrate that IL-17 directly promotes growth of lymphatic vessels by inducing increased expression of prolymphangiogenic VEGF-D and proliferation of lymphatic endothelial cells. However, IL-17-induced growth of blood vessels is primarily mediated through IL-1β secretion by IL-17-responsive cells. Furthermore, in vivo blockade of IL-17 in a preclinical model of Th17-dominant autoimmune ocular disease demonstrates a significant reduction in the corneal lymphangiogenesis and in the progression of clinical disease. Taken together, our findings demonstrate a novel prolymphangiogenic function for Th17/IL-17, indicating that IL-17 can promote the progression and amplification of immunity in part through its induction of lymphangiogenesis.

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Figures

Figure 1
Figure 1
IL-17 promotes lymphangiogenesis by inducing VEGF-D secretion, and proliferation of and tube formation by LECs. (A) Pellets containing 100 ng of IL-1β, IL-17, or IL-17 along with systemic blockade of IL-1β or VEGFR3, were placed in corneal micropockets (n = 6 mice/group) to induce angiogenesis. After 7 days, corneas were evaluated biomicroscopically and then harvested for immunostaining with CD31 (green) and Lyve1 (red). Digital micrographs using epifluorescence microscopy were captured, and ImageJ 1.34s software was used to quantify the growth of (B) blood (CD31hiLyve1−) and (C) lymphatic (CD31loLyve1+) vessels. (D) Primary human corneal epithelial cells were cultured with 10 ng/mL concentration of IL-1β, IL-17, and IL-17 with IL-1β–blocking antibodies for 24 hours, and expression levels of VEGF-A, VEGF-C, and VEGF-D mRNA in cells, and protein in culture supernatants, were measured by real-time PCR and ELISA, respectively. (E) Primary human LECs were cultured with 10 ng/mL concentrations of IL-1β, IL-17, and IL-17 with blockade of IL-1β or VEGFR-3 for 24 hours, and then proliferation was measured using BrdU incorporation assay. (F) In a transwell Matrigel assay, in vitro polarized 2 × 105 Th17 cells (in transwell) were cocultured with 5 × 104 LECs (on Matrigel) in basal MEM with reduced serum (2% FBS). Positive controls consisted of LEC cultures on Matrigel in MEM supplemented with growth factors (5% FBS, VEGF, FGF, EGF). Negative controls consisted of LEC cultures on Matrigel in basal MEM only with reduced serum (2% FBS). In some wells of Th17-LEC coculture, soluble-IL-17receptor-Fc or soluble-VEGFR3-Fc was added in media. After 8- and 24-hour incubation at 37°C, wells were visualized under bright-field inverted microscope to study the LEC tube formations on Matrigel, and digital micrographs were then captured for quantitative analysis of tube length. Data are mean ± SEM values of 3 independent experiments. *P < .05, as determined by Student t test. n.s. indicates not significant.
Figure 2
Figure 2
In vivo blockade of IL-17 inhibits inflammatory lymphangiogenesis in Th17-dominant autoimmune DED. DED was induced in wild-type C57BL6 mice by exposing them continuously to a dry air-controlled environment, which induces an autoimmune ocular surface disease. After 4 days of disease induction, mice were randomized into 3 groups receiving topically anti–IL-17 antibody, control isotype antibody, or left untreated. Progression of clinical disease was monitored by corneal fluorescein staining, a readout for clinical signs of dry eye inflammation, from day 0 to day 12, at which time corneas were harvested for immunohistochemical and real-time PCR analyses. (A) Corneas were immunostained with CD31 (green) and Lyve1 (red) antibodies. Digital micrographs using epifluorescence microscopy were captured and ImageJ software was used to (B) quantify the growth of blood (CD31hiLyve1−) and lymphatic (CD31loLyve1+) vessels. (C) Real-time PCR analysis was performed to measure the expression levels of lymphangiogenic-specific VEGF-D and VEGF-C in untreated, isotype Ab-treated, and anti–IL-17 Ab-treated DED corneas. Expression levels of VEGF-D and VEGF-C were normalized to GAPDH levels as an internal control and then with their levels in normal corneas. (D) Corneal fluorescein staining scores showing severity of clinical disease in untreated, isotype, and anti–IL-17 Ab-treated groups. Each group consists of 6 mice. Data are mean ± SEM. *P < .05, by Student t test. n.s. indicates not significant.
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References

    1. Palmer MT, Weaver CT. Autoimmunity: increasing suspects in the CD4+ T cell lineup. Nat Immunol. 2010;11(1):36–40. - PubMed
    1. Chauhan SK, El Annan J, Ecoiffier T, et al. Autoimmunity in dry eye is due to resistance of Th17 to Treg suppression. J Immunol. 2009;182(3):1247–1252. - PMC - PubMed
    1. Maniati E, Soper R, Hagemann T. Up for mischief? IL-17/Th17 in the tumour microenvironment. Oncogene. 2010;29(42):5653–5662. - PMC - PubMed
    1. Heidt S, Segundo DS, Chadha R, Wood KJ. The impact of Th17 cells on transplant rejection and the induction of tolerance. Curr Opin Organ Transplant. 2010;15(5):456–461. - PMC - PubMed
    1. Gaffen SL. Structure and signalling in the IL-17 receptor family. Nat Rev Immunol. 2009;9(8):556–567. - PMC - PubMed

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